Arabidopsis Circadian Clock RVE8 LNK1 LNK2 Interactome LC-MS

ABSTRACT: The timing of many molecular and physiological processes in plants occurs at a specific time of day. The circadian MYB-like transcription factor REVEILLE 8 (RVE8) interacts with its transcriptional coactivators NIGHT LIGHT INDUCIBLE AND CLOCK REGULATED 1 (LNK1) and LNK2 to promote the expression of evening-phased clock genes and cold tolerance factors. While genetic approaches have commonly been used to discover new connections within the clock and between other pathways, here we use affinity purification coupled with mass spectrometry to discover time-of-day-specific protein interactors of the RVE8-LNK1/2 complex. Our clock proteins are tagged with a 6X-His-3X-FLAG C-terminal (HFC) affinity tag and purified through anti-FLAG immunoprecipitation and His-tag isolation affinity purification. We also performed affinity purification-mass spectrometry of 35S::YFP-COR27 and 35S::GFP-COR28, which were immunoprecipitated with anti-GFP TRAP beads. Experiment was performed on 10-day-old seedlings grown under 12 hours light: 12 hours dark 22 degrees Celsius.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)

TISSUE(S): Plant Cell, Whole Body

SUBMITTER: Dmitri Nusinow  

LAB HEAD: Dmitri Nusinow

PROVIDER: PXD034089 | Pride | 2023-04-24

REPOSITORIES: Pride